Challenging human somatic testicular cell reassembly by protein kinase inhibition –setting up a functional in vitro test system
| Autor: | C. Brenker, Mina N. Mincheva, Stefan Schlatt, Joachim Wistuba |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Male
0301 basic medicine Cell biology Sex Differentiation medicine.drug_class Somatic cell Cell Carbazoles Morphogenesis lcsh:Medicine Cell Communication Article Indole Alkaloids 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Developmental biology Testis medicine Humans lcsh:Science Protein kinase A Protein Kinase Inhibitors Cells Cultured Sertoli Cells 030219 obstetrics & reproductive medicine Multidisciplinary lcsh:R Cell Differentiation Seminiferous Tubules Protein kinase inhibitor In vitro 030104 developmental biology medicine.anatomical_structure chemistry lcsh:Q K252a Protein Kinases Signal Transduction |
| Zdroj: | Scientific Reports, Vol 10, Iss 1, Pp 1-11 (2020) Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | Signalling pathways and cellular interactions defining initial processes of testis morphogenesis, i.e. cord formation, are poorly understood. In vitro cell-based systems modelling cord formation can be utilised as platforms to interrogate processes of tubulogenesis. We aimed at testing our established cord formation in vitro model using adult human testicular cells as a quantitative assay that can facilitate future studies on cord morphogenesis. We challenged the responsiveness of our system with a broad-spectrum protein kinase inhibitor, K252a. Cultured testicular cells were treated with various K252a concentrations under constant exposure and compound withdrawal. To quantify cell reaggregation changes, we performed computer-assisted phase-contrast image analysis of aggregate size and number. Cell reaggregation was analysed in detail by categorisation of aggregates into size groups and accounting for changes in aggregate number per size category. We found a dose-related disturbance of testicular cell reaggregation. K252a decreased aggregate size (IC50 of 203.3 nM) and reduced the large aggregate numbers. Video recordings revealed that treatment with K252a at a concentration above IC50 interfered with aggregate coalescence into cords. Short-term exposure and compound wash-out induced irreversible decrease in large aggregates. We propose our in vitro model as a functional platform to quantitatively investigate seminiferous tubulogenesis under pharmacological impact. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|